Postbiotic preparation for pelvic floor muscle rehabilitation and preparation method and application thereof

Abstract

A postbiotic preparation for pelvic floor muscle rehabilitation and a preparation method and application thereof. A strain of Lactobacillus plantarum Nice-02 is successfully obtained, and a postbiotic preparation product is successfully developed based on the strain.

What is claimed is: 
     
       1. A microbial agent, comprising an inactivated strain of  L. plantarum  Nice-02, preserved in CCTCC (China Center for Type Culture Collection) on Apr. 21, 2023 with a biological preservation number of CCTCC NO: M 2023571;
 wherein the microbial agent is prepared by a method comprising: 
 S1: activation culture: performing plate streaking on the  L. plantarum  Nice-02 into an activation culture medium to obtain a pure strain; 
 S2: primary seed solution preparation: picking and placing a single colony of the  L. plantarum  Nice-02 prepared in S1 into a first culture solution at 28 to 30° C. for stationary culture for 12 to 14 h to obtain a primary seed solution; 
 S3: secondary seed solution preparation: inoculating the primary seed solution into a second culture solution according to an inoculation amount of 0.1 to 5% for stationary culture at 30° C. to 40° C. for 10 to 12 h to obtain a secondary seed solution; 
 S4: tertiary seed solution preparation: inoculating the cultured secondary seed solution into a seeding tank containing a third culture solution under an aseptic condition at an inoculation amount of 0.5 to 5%, and performing stir culture at a stirring speed of 20 to 50 r/min, a temperature of 30° C. to 40° C., a pH value of 7.0 and a tank pressure of 0.03 to 0.06 MPa to obtain a tertiary seed solution after the culture for 10 to 12 h; 
 S5 : L. Plantarum  primary fermentation: inoculating the tertiary seed solution into a fermentation tank containing a sterilized culture solution at an inoculation amount of 0.5 to 5%, a temperature of 32° C., a stirring speed of 40 to 50 r/min to be cultured for 5 to 6 h; 
 S6 : L. plantarum  secondary fermentation: after 5 to 6 h, lowering the fermentation temperature to 30° C., accelerating the stirring speed to 50 to 70 r/min, keeping the tank pressure at 0.05 MPa and the natural pH, and performing fermentation for 3 to 4 h; 
 S7 : L. plantarum  tertiary fermentation: after 8 to 10 h, keeping the stirring speed unchanged, raising the fermentation temperature to 35° C., keeping the tank pressure at 0.05 MPa, keeping the pH value at 6.5, and performing fermentation for 2 h; 
 S8 : L. plantarum  quaternary fermentation: after 10 to 12 h, decelerating the stirring speed to 30 to 40 r/min, raising the fermentation temperature to 40° C., keeping the tank pressure at 0.05 MPa and the pH value at 6.5, and performing fermentation for 1 to 2 h to obtain a fermentation solution; and 
 S9 : L. plantarum  quinary fermentation: after 11 to 14 h, keeping the stirring speed unchanged, raising the temperature to 45° C., adding sterile lipoteichoic acid accounting for 0.05 to 0.2% of the volume of the fermentation solution into the fermentation tank, lowering the temperature to 20° C., and keeping the state for 30 min; 
 wherein the microbial agent is obtained by performing high-temperature inactivation and spray drying the fermentation solution prepared in S9. 
 
     
     
       2. The microbial agent according to  claim 1 , wherein in S1,
 the activation culture medium is prepared from 0.05 to 0.2% of casein, 0.05 to 0.2% of sialic acid, 0.3 to 1.0% of beef extracts, 0.5 to 2% of peptone, 0.05 to 0.2% of sodium acetate, 0.05 to 0.2% of potassium dihydrogen phosphate and 1.5 to 2.0% of agar powder, and a pH value is adjusted to 7.0. 
 
     
     
       3. The microbial agent according to  claim 2 , wherein the activation culture medium is prepared from 0.1% of casein, 0.1% of sialic acid, 0.7% of beef extracts, 1% of peptone, 0.1% of sodium acetate, 0.1% of potassium dihydrogen phosphate and 2.0% of agar powder, and a pH value is adjusted to 7.0. 
     
     
       4. The microbial agent according to  claim 1 , wherein in S2,
 the first culture solution is prepared from 0.05 to 0.2% of casein, 0.05 to 0.2% of sialic acid, 1 to 5% of fructooligosaccharide, 0.1 to 1% of yeast powder, 0.05 to 0.2% of sodium acetate and 0.05 to 0.2% of potassium dihydrogen phosphate, and a pH value is adjusted to 7.0. 
 
     
     
       5. The microbial agent according to  claim 4 , wherein the first culture solution is prepared from 0.1% of casein, 0.1% of sialic acid, 3% of fructooligosaccharide, 0.5% of yeast powder, 0.1% of sodium acetate and 0.1% of potassium dihydrogen phosphate, and a pH value is adjusted to 7.0. 
     
     
       6. The microbial agent according to  claim 1 , wherein in S3,
 the second culture solution is prepared from 0.1 to 1% of casein, 0.05 to 0.2% of sialic acid, 1 to 3% of fructooligosaccharide, 0.1 to 1% of yeast powder and 0.05 to 0.2% of Tween-80, and a pH value is natural. 
 
     
     
       7. The microbial agent according to  claim 6 , wherein the second culture solution is prepared from 0.5% of casein, 0.1% of sialic acid, 1.5% of fructooligosaccharide, 0.5% of yeast powder and 0.1% of Tween-80, and a pH value is natural. 
     
     
       8. The microbial agent according to  claim 1 , wherein in S4, the third culture solution is prepared from 0.5 to 5% of peptone, 1 to 5% of isomaltooligosaccharide, 0.1 to 0.5% of dipotassium phosphate, 0.5 to 5% of glycine, 0.1 to 1% of tyrosine, 0.05 to 0.2% of Tween-80 and 0.05 to 0.2% of a polyether defoamer, and a pH value is natural. 
     
     
       9. The microbial agent according to  claim 1 , wherein the third culture solution is prepared from 1% of peptone, 4% of isomaltooligosaccharide, 0.2% of dipotassium phosphate, 1% of glycine, 0.5% of tyrosine, 0.1% of Tween-80 and 0.1% of a polyether defoamer, and a pH value is natural. 
     
     
       10. The microbial agent according to  claim 1 , wherein the culture solution used in a multi-stage fermentation process is prepared from 3% of isomaltooligosaccharide, 0.2% of casein, 2% of peptone, 5% of fructooligosaccharide, 3% of water-soluble starch, 0.1% of Tween-80, 0.5% of glycine, 0.5% of tyrosine and 0.2% of a polyether defoamer, and a pH value is natural. 
     
     
       11. A method for rehabilitating pelvic floor muscles, comprising administering an amount of a microbial agent according to  claim 1  to a subject in need thereof, wherein the administration is performed orally or vaginally.